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  • A new multiplexed magnetic ...
    Tschritter, Christina M.; V. C. de Groot, Peter; Branigan, Marsha; Dyck, Markus; Sun, Zhengxin; Lougheed, Stephen C.

    Ecology and evolution, November 2023, Letnik: 13, Številka: 11
    Journal Article

    Anthropogenic stressors are exacerbating the emergence and spread of pathogens worldwide. In regions like the Arctic, where ecosystems are particularly susceptible, marked changes are predicted in regional diversity, intensity, and patterns of infectious diseases. To understand such rapidly changing host‐pathogen dynamics and mitigate the impacts of novel pathogens, we need sensitive disease surveillance tools. We developed and validated a novel multiplexed, magnetic capture, and ddPCR tool for the surveillance of multiple pathogens in polar bears, a sentinel species that is considered susceptible to climate change and other stressors with a pan‐Arctic distribution. Through sequence‐specific magnetic capture, we concentrated five target template sequences from three zoonotic bacteria (Erysipelothrix rhusiopathiae, Francisella tularensis, and Mycobacterium tuberculosis complex) and two parasitic (Toxoplasma gondii and Trichinella spp.) pathogens from large quantities (<100 g) of host tissue. We then designed and validated two multiplexed probe‐based ddPCR assays for the amplification and detection of the low‐concentration target DNA. Validations used 48 polar bear tissues (muscle and liver). We detected 14, 1, 3, 4, and 22 tissue positives for E. rhusiopathiae, F. tularensis, M. tuberculosis complex, T. gondii, and Trichinella spp., respectively. These multiplexed assays offer a rapid, specific tool for quantifying and monitoring the changing geographical and host distributions of pathogens relevant to human and animal health. Climate change is altering the distributions of many species, including pathogens and their hosts. We have developed new magnetic capture and multiplexed digital droplet PCR assays to detect the presence of three bacterial pathogens and two parasites in polar bear tissues from across the Canadian Arctic. We validate our assays and suggest how these new powerful assays can meaningfully contribute to community‐based monitoring of pathogens in polar bears and other wildlife.